Fungus Aspergillus flavus produces aflatoxin whose potency & toxicity makes them highly carcinogenic causing severe contamination to the food sources & making them primary health hazards even at low concentration. During pre & post-harvest conditions, aflatoxin contamination due to fungal infection has been reported in food grains such as groundnuts, jowar, bajra, rice, maize & wheat. Thus detection of aflatoxin concentrations in food becomes necessary. However, due to low concentration, the analytical methods applied required to be specific sensitive& simple to carry out. High- performance liquid chromatography (HPLC), was applied in detection & quantification of aflatoxin.